Ebola virus (EBOV) belongs to the Filoviridae family and causes severe illnesses such as hemorrhagic
fever with a high mortality rate up to 90%. Now two antibody drugs termed
Inmazeb and
Ebanga have been approved for treating EBOV
infection. However, clinical studies have demonstrated that the mortality rate of the patients who received these two antibody drugs remains above 30%. Therefore, novel
therapeutics with better efficacy is still desired. The isolated human
IgG1 constant domain 2 (CH2 domain) has been proposed as a scaffold for the development of C-based
single domain antibodies (C-sdAbs) as therapeutic candidates against
viral infections and other diseases. Here, we screened and identified a novel C-sdAb termed M24 that targets EBOV
glycoprotein (GP) from a C-sdAb phage display library. M24 neutralizes the pseudotype EBOV with IC50 of 0.8 nmol/L (12 ng/mL) and has modest neutralizing activity against authentic EBOV.
Epitope determination, including molecular docking and site mutation analysis, discloses that M24 binds to the internal fusion loop (IFL) within GP2, a transmembrane subunit of GP. Interestingly, we found that the binding of M24 to GP at pH 5.5 has dramatically decreased compared to the binding at pH 7.5, which may lead to weak efficacy in the neutralization of authentic EBOV. Since no sdAb against EBOV
infection has been reported to date, our results not only give a proof of concept that sdAbs could be utilized for the development of potential therapeutic candidates against EBOV
infection, but also provide useful information for the discovery and improvement of anti-EBOV agents.